Cargando…

Mechanism of SARS-CoV-2 polymerase stalling by remdesivir

Remdesivir is the only FDA-approved drug for the treatment of COVID-19 patients. The active form of remdesivir acts as a nucleoside analog and inhibits the RNA-dependent RNA polymerase (RdRp) of coronaviruses including SARS-CoV-2. Remdesivir is incorporated by the RdRp into the growing RNA product a...

Descripción completa

Detalles Bibliográficos
Autores principales: Kokic, Goran, Hillen, Hauke S., Tegunov, Dimitry, Dienemann, Christian, Seitz, Florian, Schmitzova, Jana, Farnung, Lucas, Siewert, Aaron, Höbartner, Claudia, Cramer, Patrick
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7804290/
https://www.ncbi.nlm.nih.gov/pubmed/33436624
http://dx.doi.org/10.1038/s41467-020-20542-0
_version_ 1783636130880552960
author Kokic, Goran
Hillen, Hauke S.
Tegunov, Dimitry
Dienemann, Christian
Seitz, Florian
Schmitzova, Jana
Farnung, Lucas
Siewert, Aaron
Höbartner, Claudia
Cramer, Patrick
author_facet Kokic, Goran
Hillen, Hauke S.
Tegunov, Dimitry
Dienemann, Christian
Seitz, Florian
Schmitzova, Jana
Farnung, Lucas
Siewert, Aaron
Höbartner, Claudia
Cramer, Patrick
author_sort Kokic, Goran
collection PubMed
description Remdesivir is the only FDA-approved drug for the treatment of COVID-19 patients. The active form of remdesivir acts as a nucleoside analog and inhibits the RNA-dependent RNA polymerase (RdRp) of coronaviruses including SARS-CoV-2. Remdesivir is incorporated by the RdRp into the growing RNA product and allows for addition of three more nucleotides before RNA synthesis stalls. Here we use synthetic RNA chemistry, biochemistry and cryo-electron microscopy to establish the molecular mechanism of remdesivir-induced RdRp stalling. We show that addition of the fourth nucleotide following remdesivir incorporation into the RNA product is impaired by a barrier to further RNA translocation. This translocation barrier causes retention of the RNA 3ʹ-nucleotide in the substrate-binding site of the RdRp and interferes with entry of the next nucleoside triphosphate, thereby stalling RdRp. In the structure of the remdesivir-stalled state, the 3ʹ-nucleotide of the RNA product is matched and located with the template base in the active center, and this may impair proofreading by the viral 3ʹ-exonuclease. These mechanistic insights should facilitate the quest for improved antivirals that target coronavirus replication.
format Online
Article
Text
id pubmed-7804290
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-78042902021-01-21 Mechanism of SARS-CoV-2 polymerase stalling by remdesivir Kokic, Goran Hillen, Hauke S. Tegunov, Dimitry Dienemann, Christian Seitz, Florian Schmitzova, Jana Farnung, Lucas Siewert, Aaron Höbartner, Claudia Cramer, Patrick Nat Commun Article Remdesivir is the only FDA-approved drug for the treatment of COVID-19 patients. The active form of remdesivir acts as a nucleoside analog and inhibits the RNA-dependent RNA polymerase (RdRp) of coronaviruses including SARS-CoV-2. Remdesivir is incorporated by the RdRp into the growing RNA product and allows for addition of three more nucleotides before RNA synthesis stalls. Here we use synthetic RNA chemistry, biochemistry and cryo-electron microscopy to establish the molecular mechanism of remdesivir-induced RdRp stalling. We show that addition of the fourth nucleotide following remdesivir incorporation into the RNA product is impaired by a barrier to further RNA translocation. This translocation barrier causes retention of the RNA 3ʹ-nucleotide in the substrate-binding site of the RdRp and interferes with entry of the next nucleoside triphosphate, thereby stalling RdRp. In the structure of the remdesivir-stalled state, the 3ʹ-nucleotide of the RNA product is matched and located with the template base in the active center, and this may impair proofreading by the viral 3ʹ-exonuclease. These mechanistic insights should facilitate the quest for improved antivirals that target coronavirus replication. Nature Publishing Group UK 2021-01-12 /pmc/articles/PMC7804290/ /pubmed/33436624 http://dx.doi.org/10.1038/s41467-020-20542-0 Text en © The Author(s) 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Kokic, Goran
Hillen, Hauke S.
Tegunov, Dimitry
Dienemann, Christian
Seitz, Florian
Schmitzova, Jana
Farnung, Lucas
Siewert, Aaron
Höbartner, Claudia
Cramer, Patrick
Mechanism of SARS-CoV-2 polymerase stalling by remdesivir
title Mechanism of SARS-CoV-2 polymerase stalling by remdesivir
title_full Mechanism of SARS-CoV-2 polymerase stalling by remdesivir
title_fullStr Mechanism of SARS-CoV-2 polymerase stalling by remdesivir
title_full_unstemmed Mechanism of SARS-CoV-2 polymerase stalling by remdesivir
title_short Mechanism of SARS-CoV-2 polymerase stalling by remdesivir
title_sort mechanism of sars-cov-2 polymerase stalling by remdesivir
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7804290/
https://www.ncbi.nlm.nih.gov/pubmed/33436624
http://dx.doi.org/10.1038/s41467-020-20542-0
work_keys_str_mv AT kokicgoran mechanismofsarscov2polymerasestallingbyremdesivir
AT hillenhaukes mechanismofsarscov2polymerasestallingbyremdesivir
AT tegunovdimitry mechanismofsarscov2polymerasestallingbyremdesivir
AT dienemannchristian mechanismofsarscov2polymerasestallingbyremdesivir
AT seitzflorian mechanismofsarscov2polymerasestallingbyremdesivir
AT schmitzovajana mechanismofsarscov2polymerasestallingbyremdesivir
AT farnunglucas mechanismofsarscov2polymerasestallingbyremdesivir
AT siewertaaron mechanismofsarscov2polymerasestallingbyremdesivir
AT hobartnerclaudia mechanismofsarscov2polymerasestallingbyremdesivir
AT cramerpatrick mechanismofsarscov2polymerasestallingbyremdesivir